DETERMINATION OF BENZOIC ACID AND SALICYLIC ACID

IN COMMERCIAL BENZOIC AND SALICYLIC ACIDS OINTMENTS

BY SPECTROPHOTOMETRIC METHOD
IQBAL AHMAD* AND FAIYAZ HM VAID**
*Dubai Pharmacy College, P.O. Box 19099, Dubai, United Arab Emirates
**Department of Pharmaceutical Chemistry, Faculty of Pharmacy,
University of Karachi, Karachi-75270, Pakistan

ABSTRACT
Commercial benzoic acid and salicylic acid ointments have been analyzed for benzoic acid and salicylic acid
content by using a spectrophotometric method. Since benzoic acid and salicylic acid exhibit overlapping
spectra, absorbance measurements are made at two wavelengths, i.e. 271 nm and 303 nm in ethanol (96%v/v)
for simultaneous determination of the two compounds. The method is direct and involves only one step of
absorbance measurement in the assay. The precision of the method, based on the analysis of synthetic mixtures
of the two compounds, is within 2%. The recoveries of benzoic acid and salicylic acid in the ointments range
from 99.9 to 100.6% and 99.5 to 101.3%, respectively and are comparable with those of the USP method (99.8
to 100.5% and 100.2 to 101.7%, respectively). The proposed method is simple, rapid, precise and convenient
for the assay of benzoic acid and salicylic acid in commercial preparations.

Keywords: Benzoic acid; salicylic acid; spectrophotometric assay; commercial ointments.

INTRODUCTION acid and salicylic acid in commercial ointment

The official methods for the assay of benzoic acid and
salicylic acid in compound benzoic acid ointment or
benzoic and salicylic acids ointment are based on acid-
base titration (benzoic acid) and spectrophotometric
determination (salicylic acid) (BP, 2003) or column
chromatographic separation of the two compounds
followed by spectrophotometric determination (USP,
2004). Spectrofluorimetric (Adams and Miller, 1978) and
isoabsorptive methods (Sethi, 1985) have been proposed
for the assay of benzoic acid and salicylic acid in
pharmaceutical preparations. These methods are tedious,
time consuming and may be influenced by possible
interference from related substances. Two/three-
component spectrophotometric methods have been
successfully applied to the simultaneous determination of
pharmaceutical (Ahmad et al., 1979; Ahmad et al., 1980;
Salinas et al., 1990; Santoni et al., 1990; Ahmad and
Rapson, 1990; Heelis et al., 1980; Ahmad et al., 1992;
Ahmad and Hussain, 1992; Garrido Frenich et al., 1995;
Gangwal and Sharma, 1996; Bhatia et al., 1997; Mahgoub
and Aly, 1998; Hund et al., 1999; Ribone et al., 1999;
Dinc et al., 2001; Dinc et al., 2004; Wahbi et al., 2003;
Ahmad et al., 2003; Ahmad et al., 2004a; 2004b) and
biological compounds (Zwart et al., 1984; Shih et al.,
1997).
The object of this investigation is to develop a direct,
simple, rapid, accurate, and precise spectrophotometric
method for the simultaneous determination of benzoic
preparations.
EXPERIMENTAL
Materials
Benzoic acid and salicylic acid (USP grade) were
obtained from Sigma Chemical Co., St. Louis, MO.
Ethanol (96% v/v) was analytical grade from BDH, Poole,
Dorset, UK. All chemicals and reagents were analytical
grade or of the purest form available from Sigma
Chemical Co. / BDH.
Formulation
Whitfield’s ointment containing 6% w/w benzoic acid and
3% w/w salicylic acid. Three different batches of the
ointment were used in this study.
Equipment
A Beckman Du-70 microprocessor controlled spectro-
photometer (Beckman Coulter, Fullerton, CA), was used
for absorbance measurement using 1-cm quartz cells.
Method of Analysis
Extraction of benzoic acid and salicylic acid
A portion of the ointment sample equivalent to about 60
mg of benzoic acid and 30 mg of salicylic acid was
warmed with 25 ml of ethanol (96% v/v) to melt the base
and extracted with the solvent. It was further extracted

*Present address: Baqai Institute of Pharmaceutical Sciences, Baqai Medical University, Karachi.
**Corresponding author: Tel: 9261300-7, Ext 2203 / 3203, e-mail: fvaid7@yahoo.com
18 Pak. J. Pharm. Sci., Vol.22, No.1, January 2009, pp.18-22
 Iqbal Ahmad and Faiyaz HM Vaid

Fig. 1: UV absorption spectra of (A) benzoic acid (6.8 mg/dL) and (B) salicylic acid (3.8 mg/dL) in ethanol (96%
v/v).

Fig. 2: UV absorption spectrum of ethanolic (96% v/v) extract of an ointment sample containing 0.012% w/w of
benzoic acid and 0.006% w/w salicylic acid.

with two 25 ml aliquots of ethanol (96% v/v). The
combined extracts were filtered and made up to 100 ml
with ethanol (96% v/v).
Assay of benzoic acid and salicylic acid
A 5 ml aliquot of the extract was diluted to 50 ml in a
volumetric flask with ethanol (96% v/v) and the
absorbance measurements were carried out at two
wavelength, i.e., 271 and 303 nm. The concentrations of
benzoic acid and salicylic acid were determined by a two-
component spectrophotometric assay using built-in
software.
Pak. J. Pharm. Sci., Vol.22, No.1, January 2009, pp.18-22
The assay involves the solution of two simultaneous
equations (Fell, 1986):
Aλ1 = k1λ1 C1 + k2 λ1C2 (1)
Aλ2 = k1λ2 C1 + k2 λ2 C2 (2)
Where k1λ1 and k2λ1 are the specific absorbance [A(1%, 1
cm)] values of component 1 and 2 at λ1; k1λ2 and k2λ2 are
the specific absorbance values of component 1 and 2 at
λ2; and C1 and C2 are the concentrations of component 1
and 2, respectively. The equations are solved to obtain
the concentration of each component.
19
Determination of benzoic acid and salicylic acid in commercial benzoic and salicylic acids ointments

k2λ2 Aλ1 – k2λ1Aλ2 Validity of Beer’s law

C1 = (3)
k1λ1 k2λ2 - k2λ1k1λ2
k1λ1 Aλ2 – k1λ2Aλ1
C2 = (4)
k1λ1 k2λ2 - k2λ1k1λ2
RESULTS AND DISCUSSION
Selection of analytical wavelengths
Important considerations in the selection of appropriate
wavelengths for spectrophotometric determination of
binary mixtures are maximum sensitivity (such as that at
the absorption maxima), good adherence to Beer’s law
and minimum interference from instrumental factors
(Christian, 1994). Factors influencing the selection of
analytical wavelengths in multicomponent analysis have
been studied and methods of their optimization have been
proposed (Stearns, 1969; Knowles and Burgess, 1984;
Frans and Harris, 1985; Sasaki et al., 1986; Faber et al.,
2003).
The absorption spectra of benzoic acid and salicylic acid
in ethanol (96% v/v) are sufficiently distinct to allow for
the selection of their respective absorption maxima, i.e.,
271 and 303 nm, for the analysis of the two compounds
(fig. 1). At these wavelengths maximum specificity and
sensitivity may be achieved for the analysis of these
compounds.
Table 2: Analysis of synthetic mixtures of benzoic acid and salicylic acid*
The validity of Beer’s law for benzoic acid and salicylic
acid, in the concentration range 0.2 – 1.2 × 10-2 g/dL,
alone or in mixtures, at the analytical wavelengths was
established prior to the analysis. The values of specific
absorbance [A(1%, 1 cm)] (table 1) used for the
calculation of concentrations of benzoic acid and salicylic
acid represent the means of determinations at atleast five
different concentrations (correlation coefficients 0.998 –
0.999). These values are in agreement with those
previously reported (Fell 1986; O’Neil, 2001).
Table 1: Specific absorbance [A(1%, 1 cm)] of benzoic
acid and salicylic acid in ethanol (96%, v/v)*
Compound 271 nm 303 nm
Benzoic acid 66.4 0.35
Salicylic acid 33.0 260.2
*Each value is a mean of five to seven determinations.
Reproducibility of the assay method
The reproducibility of the assay method was tested by
preparing several synthetic mixtures containing different
proportions of benzoic acid and salicylic acid and
analyzing for the two compounds by the proposed
method. The results of the assay of benzoic acid and
salicylic acid in mixtures are presented in table 2. Judging
from the calculated values of percentage recovery, the
reproducibility of the method appears to be within ± 2%.
Therefore, the method may be considered reliable and
precise for the assay of benzoic acid and salicylic acid in

Benzoic acid Salicylic acid

Added Found Recovery RSD Added Found Recovery RSD
(g × 103) (g × 103) (%) (%) (g × 103) (g × 103) (%) (%)
12.00 11.92 99.3 1.1 6.00 6.07 101.1 1.8
9.60 9.58 99.8 0.5 4.80 4.82 100.4 0.8
7.20 7.31 101.5 0.8 3.60 3.59 99.7 0.7
6.00 6.09 101.5 0.9 3.00 3.00 100.0 0.5
6.00 6.07 101.2 1.0 1.50 1.48 98.7 0.7
4.80 4.78 99.6 0.3 2.40 2.39 99.6 1.6
3.60 3.65 101.4 1.6 1.80 1.82 101.1 1.0
3.00 2.97 99.0 0.3 3.00 3.03 101.0 0.8
*Values expressed as a mean of three to five determinations.

Table 3: Assay results for benzoic acid and salicylic acid in the ointment*
Benzoic acid (6% w/w) Salicylic acid (3% w/w)
Batch
Percent of label claim (g) Percent of label claim (g)
no.
Proposed method USP method Er % Proposed method USP method Er %
1 99.9 ± 0.6 100.5 ± 1.2 - 0.6 101.3 ± 1.0 101.2 ± 1.7 + 0.1
2. 100.4 ± 1.0 101.6 ± 1.7 - 1.2 100.9 ± 1.5 101.7 ± 1.5 - 0.8
3. 100.6 ± 1.2 99.8 ± 1.5 + 0.8 99.5 ± 1.2 100.5 ± 1.9 - 1.0
*Mean ± SD, n = 3-5, Er = relative error in spectrophotometric versus USP method.

20 Pak. J. Pharm. Sci., Vol.22, No.1, January 2009, pp.18-22


commercial preparations.
Application of the method to the assay of ointments
The main object of the development of an analytical
method is to establish optimum assay conditions to
achieve a high degree of specificity, sensitivity and
precision and its validation (Green, 1996). This has been
achieved in the present study. The proposed method has
been validated and applied to the assay of benzoic acid
and salicylic acid in ointment preparations. The UV
absorption spectrum of the ethanolic extract of an
ointment sample is shown in fig. 2. The spectrum exhibits
two absorption maxima at about 271 nm and 303 nm
corresponding to a mixture of benzoic acid and salicylic
acid which can be resolved by a two-component assay.
The assay results obtained by the proposed method for
benzoic acid and salicylic acid in three different batches
of the ointment along with those obtained by the USP
method are presented in table 3. The recoveries of benzoic
acid and salicylic acid in the samples assayed by the
proposed method are in good agreement with those of the
USP method and the relative error of the proposed
method versus USP method is within ± 1%. The method
has the advantage of being simple, rapid and convenient
involving only one step of absorbance measurements in
the assay of benzoic acid and salicylic acid in commercial
ointments.
CONCLUSION
A two-component spectrophotometric method has been
developed for the direct simultaneous determination of
benzoic acid and salicylic acid in commercial ointment
preparations. The method is simple, rapid, accurate and
precise for the assay of benzoic acid and salicylic acid. It
is convenient for the determination of the two compounds
in pharmaceutical preparations in the quality control
laboratories.
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